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系統識別號 U0026-3107201523430700
論文名稱(中文) 研究糖尿病病患幽門桿菌除菌率偏低的機轉:高葡萄糖濃度引發clarithromycin抗藥性或加速clarithromycin降解
論文名稱(英文) Investigate the mechanism for poor eradication rate of Helicobacter pylori in diabetic patients
校院名稱 成功大學
系所名稱(中) 臨床醫學研究所
系所名稱(英) Institute of Clinical Medicine
學年度 103
學期 2
出版年 104
研究生(中文) 吳建霆
研究生(英文) Chien-Ting Wu
學號 S97011064
學位類別 碩士
語文別 中文
論文頁數 45頁
口試委員 指導教授-楊燿榮
口試委員-許博翔
口試委員-楊曉白
口試委員-張維倫
中文關鍵字 葡萄糖  幽門桿菌  clarithromycin抗藥性  clarithromycin降解 
英文關鍵字 glucose  Helicobacter pylori  clarithromycin  resistance  degradation 
學科別分類
中文摘要 幽門桿菌感染是全世界最盛行的細菌感染疾病之一。幽門桿菌會造成慢性胃炎、潰瘍性疾病、甚至胃癌的發生,因此根除幽門桿菌感染就顯得相當重要。目前,以氫離子幫浦阻斷劑、clarithromycin、amoxicillin治療7至14天的三合治療,仍是常用的第一線處方。在台灣,三合治療的成功除菌率大約是82-94%。然而,研究顯示糖尿病病患與非糖尿病病患相比,有較高的幽門桿菌除菌失敗風險。其中,又以clarithromycin為基底的處方更為明顯。在糖尿病病患,以clarithromycin為基底的三合治療及序列治療成功除菌率只有約50%。糖尿病病患幽門桿菌除菌率較差的原因仍不明確。本研究發現之前曾經除菌失敗過的糖尿病病患比非糖尿病病患,有較高的clarithromycin抗藥性。而且糖尿病病患胃液中的葡萄糖濃度也較非糖尿病病患的稍高。過去研究顯示葡萄糖對於細菌及黴菌的抗藥性有所影響,也會對抗生素的穩定性有所影響。因此,我設計了實驗要探討葡萄糖在體外是否會引發幽門桿菌對clarithromycin的抗藥性及是否會影響clarithromycin的降解。
第一個體外實驗,我選取了具有clarithromycin敏感性的幽門桿菌標準菌株J99,以及從成大醫院糖尿病病患胃中分離出來的臨床菌株。我將這些菌株置於布魯氏菌培養肉湯中,並額外添加葡萄糖及低於致死劑量的clarithromycin (葡萄糖濃度分別為:100、200及300 mg/dl;clarithromycin濃度分別為:0.015及0.031 μg/ml),用振盪器於37℃微需氧環境中培養。初代培養三天後,更換一次培養液,進行繼代培養,直到第27天。接著於第三、六、九代利用瓊脂稀釋法測試幽門桿菌的最小抑菌濃度。最小抑菌濃度大於等於1 μg/ml即是對clarithromycin有抗藥性。並以L-form葡萄糖當作osmolality control。
第二個體外實驗,我抽取來成大醫院做胃鏡的病人的胃液,離心去除黏液後,利用pH meter測定胃液的pH值,利用glucose assay kit測定胃液中的葡萄糖濃度。接著我將clarithromycin配製在不同葡萄糖濃度的胃液和水溶液中 (葡萄糖濃度分別為:0、100、200及300 mg/dl),放入37℃的培養箱。每24小時收集一次溶液樣本,立即冷凍於-20℃的冰箱中,連續收集七天。最後利用高效液相層析法分析溶液樣本中clarithromycin的剩餘濃度,來計算其降解的情況。
本研究發現在沒有clarithromycin的培養肉湯中,幽門桿菌菌株在不同葡萄糖濃度下,生長狀況良好。然而,不論是標準菌株J99或從糖尿病病人胃中分離出來的臨床菌株 (包含三株clarithromycin-sensitive菌株、兩株clarithromycin-intermediate菌株、及一株clarithromycin-resistant菌株),葡萄糖均不會使其最小抑菌濃度上升,意即在本研究的設計下,葡萄糖不會促使幽門桿菌產生抗藥性菌株。另一部分,配製在不同葡萄糖濃度的胃液和水溶液中的clarithromycin,直到第七天,這些溶液樣本中clarithromycin的剩餘濃度並無顯著減低,意即葡萄糖不會加速clarithromycin的降解。
因此,本研究推論造成糖尿病病患幽門桿菌除菌率低的原因,可能不是因為高葡萄糖濃度引發幽門桿菌對clarithromycin的抗藥性,也不是高葡萄糖濃度加快clarithromycin的降解。
英文摘要 The eradication rate of Helicobacter pylori (H. pylori) in the patients with diabetes mellitus (DM) was poor. The mechanism remained unclear. I found that higher glucose levels neither induced the clarithromycin-resistant H. pylori nor enhanced clarithromycin degradation, which may correspond to eradication failure of H. pylori in diabetic patients.
論文目次 中文摘要 I
英文延伸摘要 IV
誌謝 IX
目錄 X
內文目錄 X
表目錄 XIII
圖目錄 XIV
符號及縮寫對照表 XV
第一章 緒論 1
一、幽門桿菌介紹 1
二、幽門桿菌與代謝性疾病的關係 1
三、糖尿病病患的幽門桿菌除菌率 2
四、糖尿病病患幽門桿菌除菌率低的因素 3
五、研究目標 5
第二章 研究設計及方法 6
一、糖尿病病患是否有較高的幽門桿菌抗藥性 6
二、高葡萄糖濃度是否會升高幽門桿菌對clarithromycin的MICs 6
(一) 幽門桿菌培養 6
(二) 葡萄糖引發幽門桿菌抗藥性實驗 7
(三) 利用瓊脂稀釋法測試最小抑菌濃度 7
三、高葡萄糖濃度是否會加快clarithromycin的降解 8
(一) 胃液收集及化學分析 8
(二) 利用高效液相層析法測定clarithromycin的剩餘濃度 8
1. 高效液相層析法的實驗條件 9
2. Clarithromycin在不同葡萄糖濃度水溶液中的剩餘濃 9
3. Clarithromycin在不同葡萄糖濃度胃液中的剩餘濃度 10
(三) 資料分析 10
第三章 結果 11
一、糖尿病病患幽門桿菌抗藥性比率 11
二、不同葡萄糖濃度中幽門桿菌的生長情況 12
三、高葡萄糖濃度對幽門桿菌抗藥性的影響 13
四、糖尿病病患胃液中葡萄糖濃度及pH值 14
五、高葡萄糖濃度對clarithromycin降解的影響 15
第四章 討論 17
參考文獻 23
附錄 33
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